Identification of oxidation products of (-)-epigallocatechin gallate and (-)-epigallocatechin with H(2)O(2)

(-)-Epigallocatechin gallate (EGCG) and (-)-epigallocatechin (EGC) are two important antioxidants in tea. They also display some antitumor activities, and these activities are believed to be mainly due to their antioxidative effects. However, the specific mechanisms of antioxidant action of tea catechins remain unclear. In this study are isolated and identified two novel reaction products of EGCG and one product of EGC when they were reacted separately with H(2)O(2). These products are formed by the oxidation and decarboxylation of the A ring in the catechin molecule. This study provides unequivocal proof that the A ring of EGCG and EGC may also be an antioxidant site. This study also indicates an additional reaction pathway for the oxidation chemistry of tea catechins.     

Excretion of (14)C-fumonisin B(1), (14)C-hydrolyzed fumonisin B(1), and (14)C-fumonisin B(1)-fructose in rats.

14C-Fumonisin B(1) (FB(1)) was produced by Fusarium proliferatum M-5991 in modified Myro liquid medium and purified to >95% purity with a specific activity of 1.7 mCi/mmol. Nine male and nine female F344/N rats were each dosed by gavage with 0.69 micromol of (14)C-FB(1), (14)C-hydrolyzed FB(1), or (14)C-FB(1)-fructose/kg body weight. Urinary excretion of (14)C-FB(1) and (14)C-FB(1)-fructose was 0.5% and 4.4% of the total dose, respectively, and was similar between male and female rats. Urinary excretion of (14)C-hydrolyzed HFB(1) was significantly greater (P > 0.05) in female rats as compared with male rats (17.3% vs 12.8% of the total dose, respectively). There were no significant (P > 0.05) differences in biliary excretion of the three fumonisin compounds with a mean of 1. 4% of the dose excreted at 4 h after dosing. Lesser amounts continued to be excreted up to 9.25 h after dosing. Although biliary excretion of the (14)C-FB(1), (14)C-hydrolyzed FB(1), and (14)C-FB(1)-fructose was similar, increased urinary excretion of the (14)C-hydrolyzed FB(1) as compared to (14)C-FB(1) and (14)C-FB(1)-fructose indicated a greater absorption of the hydrolyzed form.     

Chemoenzymatic synthesis of homochiral (R)- and (S)-karahanaenol from (R)-limonene

Terpinolene oxide, a monoterpene belonging to the p-menthane group, is easily derived from naturally abundant (R)-limonene. It was isomerized with montmorillonite clay catalyst to karahanaenone (2,2, 5-trimethylcyclohept-4-en-1-one) by ring enlargement. The enantiomers of the corresponding alcohol, karahanaenol (2,2, 5-trimethylcyclohept-4-en-1- ol), known for their individual organoleptic properties, were resolved through Pseudomonas cepacia lipase mediated enantiospecific alcoholysis of its acetate derivative.     

(三唑基-~(14)C-)粉锈宁的标记合成

本文报道了(三唑基-14C)-粉锈宁的制备。由14C-甲酸和重碳酸氨基胍形成(5-14C)-3-氨基-1,2,4-三唑,再经重氮化脱氨得到(5-14C)-1,2,4-三唑,最后再与对氯酚和二氯片呐酮反应得到(三唑基-14C)-粉锈宁。放化收率为26％(从甲酸-14C计),放化纯度大于95％。     

Theoretical speciation of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) in agronomic conditions

The presence of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) as the second largest component in commercial EDDHA iron chelates has recently been demonstrated. Here is reported the speciation of o,p-EDDHA by the application of a novel methodology through the determination of the complexing capacity, protonation, and Ca(2+), Mg(2+), Cu(2+), and Fe(3+) stability constants. The pM values and species distribution in solution, hydroponic, and soil conditions were obtained. Due to the para position of one phenol group in o,p-EDDHA, the protonation constants and Ca and Mg stability constants have different values from those of o,o-EDDHA and p,p-EDDHA regioisomers. o,p-EDDHA/Fe(3+) stability constants are higher than those of EDTA/Fe(3+) but lower than those of o,o-EDDHA/Fe(3+). The sequence obtained for pFe is o,o-EDDHA/Fe(3+) >/= o,p-EDDHA/Fe(3+) > EDTA/Fe(3+). o,p-EDDHA/Fe(3+) can be used as an iron chelate in hydroponic conditions. Also, it can be used in soils with limited Cu availability.     

Binding studies of oxovanadium(V) with ovalbumins (OA)

The effect of protein oxovanadium(V) ion concentration and pH on the ratio of diffusion current (id/id₀) was studied in vanadium(V) ovalbumin-S and denatured ovalbumin systems. In both the cases marked decrease in diffusion current was observed at the respective pH values, indicating that binding takes place with cationic groups of the proteins. The binding sites (n) were found to be pH dependent. The uniformity of logK and ΔG ⁰ value at all pH values indicated the involvement of same sites in interaction. Furthermore, the linear scatchard plots in both the systems supported the involvement of single class of independent sites in oxovanadium(V) anion interaction. The difference in binding sites (n) has been attributed to the folded structure of ovalbumin-S while unfolded one of denatured ovalbumin.     

Effect of pH on interaction between As(III) or As(V) and manganese(IV) oxide

The efficiency of As(III) oxidation by MnO₂, and retention of oxidation products varies with system pH. Maximum retention by hydrous Mn(IV) oxide occurs at pH < 5, declining at higher pH to about half total As at pH 10. The adsorption capacities of pyrolusite and cryptomelane at pH ～6.5 for As(V) species were 10 and 25 mmol kg^?1, respectively. HMO surface saturation (～10 mmol kg^?1) was reached with equilibrium As(V) levels of 5 to 8 × 10^?6 M but this was supplemented at higher levels by an absorption process where uptake increased linearly with concentration (e.g., 68 mmol kg^?1 with 2 × 10^?5 M As(V)). Added As(III) was avidly oxidized and most product retained at pH 3. At higher pH increasing amounts of As(III) remained unoxidized due to initial reactions apparently blocking access to internal pores. Added Na⁺ reduced the amount of As retained by the HMO, with the phosphate salt having a significant effect. Extraction studies confirmed that most As could be released by exposure to reducing agents or chelating agents (EDTA). The environmental significance of the results has been considered.     

Quantitation of (R)- and (S)-linalool in beer using solid phase microextraction (SPME) in combination with a stable isotope dilution assay (SIDA)

A stable isotope dilution assay (SIDA) was developed for the quantitation of both linalool enantiomers using synthesized [2H(2)]R/S-linalool as the internal standard. For enrichment of the target compound from beer, a solid phase microextraction method (SPME) was developed. In comparison to the more time-consuming extraction/distillation cleanup of the beer samples, the results obtained by SPME/SIDA were very similar, even under nonequilibration conditions. Analysis of five different types of beer showed significant differences in the linalool concentrations, which were clearly correlated with the intensity of the hoppy aroma note as evaluated by a sensory panel. In addition, significant differences in the R/S ratios were measured in the beers. The SPME/SIDA yielded exact data independently from headspace sampling parameters, such as exposure time or ionic strength of the solution.     

锌指核酸酶(ZFN)介导的牛肌肉抑制素基因(MSTN)突变的研究

肌肉生成抑制素(myostatin,MSTN)是肌肉发育的负调控基因,突变后会引起肌肉的过度发育,在肉用动物育种中有非常重要的价值。本研究利用锌指核酸酶(zinc finger nuclease,ZFN)对牛(Bos taurus)MSTN基因的第二外显子特异性剪切,诱导该基因的突变,从而达到敲除该基因的目的。首先对脂质体介导的双质粒共转染牛成纤维细胞的条件进行优化,获得的共转染效率高达19.27%;之后利用两组ZFNs对牛成纤维细胞进行转染,单细胞通过口吸法移入96孔板,经扩大培养后进行PCR筛选,获得18株MSTN基因突变细胞株,其中包括一株双等位基因敲除的细胞系,两组ZFNs对牛成纤维细胞的突变效率分别为6.05%和3.84%;最后,挑选一株突变细胞系进行体细胞核移植研究,共获得24枚重构胚;将去除透明带的重构胚培养于2i培养液中,获得囊胚来源的类滋养层干细胞,通过PCR和测序检测,证明该细胞系在MSTN基因第二外显子具有与供体细胞相同的突变。结果表明,所合成的两组ZFNs可以在MSTN作用位点引起突变,具有很高的突变效率,而且能获得双等位基因突变的细胞株,获得的细胞株能够用作体细胞核移植的供体细胞。本研究为肉用牛的品质改良和育种工作提供了基础资料。     

Competition from Cu(II), Zn(II) and Cd(II) in Pb(II) Binding to Suwannee River Fulvic Acid

This is a study of trace metal competition in the complexation of Pb(II) by well-characterized humic substances, namely Suwannee River Fulvic Acid (SRFA) in model solutions. It was found that Cu(II) seems to compete with Pb(II) for strong binding sites of SRFA when present at the same concentration as Pb(II). However, Cd(II) and Zn(II) did not seem to compete with Pb(II) for strong binding sites of SRFA. These two metals did compete with Pb(II) for the weaker binding sites of SRFA. Heterogeneity of SRFA was found to play a crucial role in metal–SRFA interactions. The environmental significance of this research for freshwater is that even at relatively low Pb(II) loadings, the metals associated with lead in minerals, e.g. Cu(II), may successfully compete with Pb(II) for the same binding sites of the naturally occurring organic complexants, with the result that some of the Pb(II) may exist as free Pb²⁺ ions, which has been reported to be one of the toxic forms of Pb in aquatic environment.     

Biotransformation of (R)- and (S)-terpinen-4-ol by the larvae of common cutworm (Spodoptera litura)

(R)-Terpinen-4-ol was mixed in an artificial diet at a concentration of 1 mg/g of diet, and the diet was fed to the last instar larvae of common cutworm (Spodoptera litura). Metabolites were recovered from frass and analyzed spectroscopically. (R)-Terpinen-4-ol was transformed mainly to (R)-p-menth-1-en-4,7-diol. Similarly, (S)-terpinen-4-ol was transformed mainly to (S)-p-menth-1-en-4,7-diol. The C-7 position (allylic methyl group) of (R)- and (S)-terpinen-4-ol was preferentially oxidized.     

Standard potentials (Eo) of iron(III) oxides under reducing conditions

Partial reduction of iron(III) oxides with hydrogen in the presence of a platinum catalyst leads to an equilibrium state after 4–20 h. From the measured Eh, pH, and Fe²⁺ concentration conditional standard potentials can be calculated using the formula E_o (volt) = Eh + 0.059 lg(Fe²⁺) + 0.18 pH which indicate the stability of Fe oxides against reduction. The reduceability decreases following the order ferrihydrite > lepidocrocite > hematite > goethite. The difference between hematite and goethite was more pronounced than that predicted from thermodynamic data.     

Anthocyanins from maize (Zea mays) and reed canarygrass (Phalaris arundinacea)

Flowers of maize, Zea mays, and reed canarygrass, Phalaris arundinacea, contain the same anthocyanins: cyanidin 3-glucoside, cyanidin 3-(6' '-malonylglucoside), cyanidin 3-(3' ',6' '-dimalonylglucoside), peonidin 3-glucoside, peonidin 3-(6' '-malonylglucoside), and peonidin 3-(dimalonylglucoside). The latter pigment has previously not been reported to occur in plants. Structure elucidations were primarily based on homo- and heteronuclear two-dimensional NMR and electrospray MS. During the isolation procedure using various mixtures of H(2)O, CF(3)CO(2)H, and CH(3)OH, and during storage in NMR solvent (CF(3)CO(2)D/CD(3)OD; 1:19, v/v) methyl esterification of the free acid function of the malonyl units of the pigments occurs. The acylated anthocyanins constitute more than 80% and 40% of the anthocyanins in P. arundinacea and Z. mays, respectively. Flowers and leaves of maize, Zea mays, contain the same anthocyanins in nearly equal relative proportions.